目的:分析HOXD10基因启动子区甲基化水平与基因表达水平的关联并探索HOXD10与肺腺癌临床病理特征和预后的关系。方法:采用R语言拓展包TCGAbiolinks进行基因差异表达分析和差异甲基化分析,利用肿瘤基因组计划(The Cancer Genome Atlas,TC...目的:分析HOXD10基因启动子区甲基化水平与基因表达水平的关联并探索HOXD10与肺腺癌临床病理特征和预后的关系。方法:采用R语言拓展包TCGAbiolinks进行基因差异表达分析和差异甲基化分析,利用肿瘤基因组计划(The Cancer Genome Atlas,TCGA)项目肺腺癌(lung adenocarcinoma,LUAD)数据集中521例癌症组织和58例癌旁组织的RNA-seq数据分析HOXD10基因差异表达,利用同一数据集中461例癌症组织和32例癌旁组织的甲基化芯片数据分析HOXD10启动子区差异甲基化,并计算启动子区差异甲基化与基因表达水平的相关性,最后分析HOXD10表达与LUAD临床病理特征和预后的关系。结果:在TCGA项目LUAD数据集中,521例癌症组织HOXD10基因的平均表达量为30.4(normalized read counts),低于58例癌旁组织的平均表达量170.8,差异具有统计学意义(P<0.05);在癌症和癌旁组织配对(即来源于同一患者的癌症和癌旁组织组成配对,n=56)时HOXD10基因表达水平的平均差异值为−141.2,差异具有统计学意义(P<0.05)。通过比较461例癌症组织和32例癌旁组织的甲基化芯片数据,鉴定出一个位于HOXD10基因启动子区内的差异甲基化区域,甲基化水平癌症组织高于癌旁组织(Δβ=0.3656,fwer=0)。该区域位于以hg19作为参考基因组的2号染色体176980837~176982342坐标位置,共包含12个甲基化探针,其中cg03918304,cg10364040,cg13217260,cg20649017,cg21591742和cg25371634等6个探针的β值与HOXD10基因的表达情况呈负相关(r<−0.1,P<0.05)。250例肺腺癌患者的完整临床信息显示:HOXD10表达水平与每天抽烟根数和肿瘤大小两个临床病理指标有关(P<0.05),HOXD10低表达组的5年中位生存时间为37.2个月,低于高表达组的59.9个月(P<0.05)。结论:HOXD10基因在TCGA肺腺癌癌症组织中的表达水平明显高于癌旁组织,而甲基化水平明显低于癌旁组织;HOXD10的表达与肺腺癌的临床病理特征有关,是肺腺癌预后的可能影响因素。展开更多
Glioblastoma is the most aggressive and invasive brain tumor and has a poor prognosis;elucidating the underlying molecular mechanisms is essential to select molecular targeted therapies.Here,we investigated the effect...Glioblastoma is the most aggressive and invasive brain tumor and has a poor prognosis;elucidating the underlying molecular mechanisms is essential to select molecular targeted therapies.Here,we investigated the effect of microRNAs on the marked invasiveness of glioblastoma.U373 glioblastoma cells were infected with 140 different microRNAs from an OncomiR library,and the effects of the invasion-related microRNAs and targeted molecules were investigated after repeated Matrigel invasion assays.Screening of the OncomiR library identified miR-23a as a key regulator of glioblastoma invasion.In six glioblastoma cell lines,a positive correlation was detected between the expression levels of miR-23a and invasiveness.A luciferase reporter assay demonstrated that homeobox D10(HOXD10)was a miR-23a-target molecule,which was verified by high scores from both the PicTar and miRanda algorithms.Forced expression of miR-23a induced expression of invasion-related molecules,including uPAR,RhoA,and RhoC,and altered expression of glial-mesenchymal transition markers such as Snail,Slug,MMP2,MMP9,MMP14,and E-cadherin;however,these changes in expression levels were reversed by HOXD10 overexpression.Thus,miR-23a significantly promoted invasion of glioblastoma cells with polarized formation of focal adhesions,while exogenous HOXD10 overexpression reversed these phenomena.Here,we identify miR-23a-regulated HOXD10 as a pivotal regulator of invasion in glioblastoma,providing a novel mechanism for the aggressive invasiveness of this tumor and providing insight into potential therapeutic targets.展开更多
基金This work was supported,in part,by Grants-in-Aid from the Ministry of Education,Culture,Sports,Science,and TechnologyJapanese Society for the Promotion of Science+1 种基金Ministry of Health,Labor,and Welfare of Japan as well as a grant from the Japanese Science and Technology AgencyIn addition,this research was supported by Global Station for Soft Matter,a project of Global Institution for Collaborative Research and Education at Hokkaido University.Institute for Chemical Reaction Design and Discovery(ICReDD)was established by World Premier International Research Initiative(WPI),MEXT,Japan.
文摘Glioblastoma is the most aggressive and invasive brain tumor and has a poor prognosis;elucidating the underlying molecular mechanisms is essential to select molecular targeted therapies.Here,we investigated the effect of microRNAs on the marked invasiveness of glioblastoma.U373 glioblastoma cells were infected with 140 different microRNAs from an OncomiR library,and the effects of the invasion-related microRNAs and targeted molecules were investigated after repeated Matrigel invasion assays.Screening of the OncomiR library identified miR-23a as a key regulator of glioblastoma invasion.In six glioblastoma cell lines,a positive correlation was detected between the expression levels of miR-23a and invasiveness.A luciferase reporter assay demonstrated that homeobox D10(HOXD10)was a miR-23a-target molecule,which was verified by high scores from both the PicTar and miRanda algorithms.Forced expression of miR-23a induced expression of invasion-related molecules,including uPAR,RhoA,and RhoC,and altered expression of glial-mesenchymal transition markers such as Snail,Slug,MMP2,MMP9,MMP14,and E-cadherin;however,these changes in expression levels were reversed by HOXD10 overexpression.Thus,miR-23a significantly promoted invasion of glioblastoma cells with polarized formation of focal adhesions,while exogenous HOXD10 overexpression reversed these phenomena.Here,we identify miR-23a-regulated HOXD10 as a pivotal regulator of invasion in glioblastoma,providing a novel mechanism for the aggressive invasiveness of this tumor and providing insight into potential therapeutic targets.
